Compositions and methods for increasing the viability of freeze-dried probiotics

Abstract

The present disclosure relates to compositions, methods, and kits for increasing the viability of bacteria that have been subjected to freeze-drying/lyophilization. In particular, the disclosure relates to compositions and methods for increasing the viability of living medicines (e.g., probiotics) during lyophilization, oral rehydration, and passage through the gastrointestinal tract

1 . A composition comprising a probiotic bacteria population and one or more tardigrade-specific intrinsically disordered proteins (TSIDPs) 
     
     
         2 . The composition of  claim 1 , wherein bacteria of the probiotic bacteria population express the one or more TSIDPs. 
     
     
         3 . The composition of  claim 1 , wherein the probiotic bacteria population comprises bacteria of one or more species selected from the genera consisting of  Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, Escherichia , and  Saccharomyces.    
     
     
         4 . The composition of  claim 3 , wherein the one or more probiotic bacterial species comprise a  Lactobacillus  or  Bifidobacterium  species. 
     
     
         5 . The composition of  claim 1 , wherein the TSIDPs are selected from the group consisting of tardigrade protein 77580, tardigrade protein 89226, tardigrade protein 94063, tardigrade protein 106094, tardigrade protein 107838, and combinations thereof. 
     
     
         6 . The composition of  claim 1 , wherein the probiotic bacteria population and the one or more TSIDPs are present in an admixture. 
     
     
         7 . A composition selected from the group consisting of:
 A preparation for oral delivery to a subject, the preparation comprising one or more recombinant probiotic bacteria that express one or more tardigrade-specific intrinsically disordered proteins (TSIDPs);   A preparation for oral delivery possessing freeze-dried recombinant probiotic bacteria;   A freeze-dried preparation for oral delivery comprising one or more isolated tardigrade-specific intrinsically disordered protein (TSIDPs) and one or more isolated populations of probiotic bacteria;   A pharmaceutical composition comprising an isolated tardigrade-specific intrinsically disordered protein (TSIDP), an isolated population of probiotic bacteria, and a pharmaceutically acceptable carrier;   A pharmaceutical composition comprising recombinant probiotic bacteria that express a tardigrade-specific intrinsically disordered protein (TSIDP), and a pharmaceutically acceptable carrier;   A composition comprising a probiotic bacteria population and one or more natural or synthetic intrinsically disordered proteins (IDPs) that possesses cryoprotective or vitrification properties;   A preparation for oral delivery to a subject, the preparation comprising one or more recombinant probiotic bacteria that express one or more intrinsically disordered proteins (IDPs);   A preparation for oral delivery comprising freeze-dried recombinant probiotic bacteria;   A freeze-dried preparation for oral delivery comprising one or more isolated intrinsically disordered protein (IDPs) and one or more isolated populations of probiotic bacteria;   A pharmaceutical composition comprising an isolated intrinsically disordered protein (IDP), an isolated population of probiotic bacteria, and a pharmaceutically acceptable carrier; and   A pharmaceutical composition comprising recombinant probiotic bacteria that express an intrinsically disordered protein (IDP), and a pharmaceutically acceptable carrier.   
     
     
         8 . The composition of  claim 7 , wherein the one or more recombinant probiotic bacteria express the one or more TSIDPs. 
     
     
         9 . The composition of  claim 7 , wherein the one or more recombinant probiotic bacteria are of species selected from the genera consisting of  Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, Escherichia , and  Saccharomyces.    
     
     
         10 . The composition of  claim 9 , wherein the one or more probiotic bacterial species comprise a  Lactobacillus  or  Bifidobacterium  species. 
     
     
         11 . The composition of  claim 9 , wherein the TSIDPs are selected from the group consisting of tardigrade protein 77580, tardigrade protein 89226, tardigrade protein 94063, tardigrade protein 106094, tardigrade protein 107838, and combinations thereof. 
     
     
         12 . (canceled) 
     
     
         13 . The composition of  claim 7 , wherein the recombinant probiotic bacteria express one or more TSIDPs selected from the group consisting of tardigrade protein 77580, tardigrade protein 89226, tardigrade protein 94063, tardigrade protein 106094, tardigrade protein 107838, and combinations thereof. 
     
     
         14 . The composition of  claim 7 , wherein the recombinant probiotic bacteria express one or more IDPs selected from the group consisting of prokaryotic and eukaryotic proteins that possess cryoprotective or vitrification properties, and combinations thereof. 
     
     
         15 . The composition of  claim 7 , wherein the one or more isolated populations of probiotic bacteria include species selected from the genera consisting of  Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, Escherichia , and  Saccharomyces.    
     
     
         16 . The composition of  claim 7 , wherein:
 bacteria of the probiotic bacteria population express the one or more IDPs;   the probiotic bacteria population comprises bacteria of one or more species selected from the genera consisting of  Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, Escherichia , and  Saccharomyces , optionally wherein the one or more probiotic bacterial species comprise a  Lactobacillus  or  Bifidobacterium  species;   the IDPs are selected from the group consisting of prokarvotic or eukarvotic proteins that possess cryoprotective or vitrification properties; and/or   the probiotic bacteria population and the one or more IDPs are present in an admixture.   
     
     
         17 . The composition of  claim 7 , wherein:
 the one or more IDPs possess cryoprotective or vitrification properties; and/or   the one or more recombinant probiotic bacteria are of species selected from the genera consisting of  Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, Escherichia , and  Saccharomyces , optionally wherein the one or more probiotic bacterial species comprise a  Lactobacillus  or  Bifidobacterium  species and/or the IDPs are selected from the group consisting of prokaryotic or eukaryotic proteins that possess cryoprotective or vitrification properties, and combinations thereof.   
     
     
         18 . A method selected from the group consisting of:
 A method for treating or preventing a probiotic-treatable disease or disorder in a subject having or at risk of developing the probiotic-treatable disease or disorder, the method comprising administering to the subject:   (a) a pharmaceutical composition comprising an isolated TSIDP, an isolated population of probiotic bacteria, and a pharmaceutically acceptable carrier, or   (b) a pharmaceutical composition comprising recombinant probiotic bacteria that express a TSIDP, and a pharmaceutically acceptable carrier; and   A method for treating or preventing a probiotic-treatable disease or disorder in a subject having or at risk of developing the probiotic-treatable disease or disorder, the method comprising administering to the subject:   (a) a pharmaceutical composition comprising an isolated IDP, an isolated population of probiotic bacteria, and a pharmaceutically acceptable carrier, or   (b) a pharmaceutical composition comprising recombinant probiotic bacteria that express an IDP, and a pharmaceutically acceptable carrier.   
     
     
         19 . The method of  claim 18 , wherein the probiotic-treatable disease or disorder is selected from the group consisting of a urea cycle disorder (UCD), phenylketonuria (PKU), inflammatory bowel disease (IBD) and a cancer. 
     
     
         20 - 38 . (canceled)